• Title/Summary/Keyword: Fluorescence analysis

Search Result 1,254, Processing Time 0.037 seconds

Repetition of Apoptosis Induced by Amiloride Derivatives in Human Umbilical Vein Endothelial Cells (제대정맥 내피세포에서 Amiloride 유도체에 의한 Apoptosis 반복)

  • Park, Kyu Chang;Park, Kyu Sang;Moon, Soo Jee
    • Clinical and Experimental Pediatrics
    • /
    • v.46 no.1
    • /
    • pp.56-66
    • /
    • 2003
  • Purpose : Human umbilical vein endothelial cells(HUVECs) play an important role in regulating blood flow by releasing vasoactive substances. It has been reported that endothelial impairment and dysfunction might be a primary cause of placental vascular disease, which is manifested clinically as preeclampsia in mother and intrauterine growth restriction in fetus. Furthermore, the frequency of apoptotic changes is increased in umbilical and placental tissues from growth-restricted pregnancies. However, the various mechanisms of umbilical endothelial cell apoptosis have not been broadly proposed. We investigate the effects of amiloride derivatives on apoptotic death of HUVECs and identify their ionic mechanism. Methods : HUVECs were purchased from Clonetics, and cultured on endothelial cell growth medium. MTT assay and flow cytometry were used for assessing cytotoxic effect and confirming the apoptosis. Changes in intracellular ion concentrations were measured with specific fluorescent dyes and fluorescence imaging analysis system. Results : Amiloride derivatives elicited cytotoxic effects on HUVECs with dose-dependent manners and the rank order of potency is HMA($IC_{50}\;11.2{\mu}M$), MIA>EIPA>>amiloride. HMA-induced cytotoxicity is dependent on extra- and intracellular pH, that is, increase extra- and intracellular pH augmented the cytotoxic effects of HMA. HMA dose-dependently reduced intracellular major ions, such as $K^+$ and $Cl^-$. Interestingly, the depletion of intracellular ions induced by HMA was also significantly enhanced at alkaline extracellular pH. Conclusion : Amiloride derivatives induce apoptosis of HUVECs with dose and pH dependent manners. They reduce intracellular $K^+$ and $Cl^-$ concentration, which is also extracellular pH dependent.

Analytical Methods of Hydroxyl Radical Produced by TiO2 Photo-catalytic Oxidation (TiO2 광촉매 산화 반응에서 생성된 수산기 라디칼 분석 방법)

  • Kim, Seong Hee;Lee, Sang-Woo;Kim, Jeong Jin;Kim, Soon-Oh
    • Journal of the Mineralogical Society of Korea
    • /
    • v.28 no.3
    • /
    • pp.245-253
    • /
    • 2015
  • The performance of $TiO_2$ photo-catalytic oxidation process is significantly dependent on the amount of hydroxyl radicals produced during the process, and it is an essential prerequisite to quantify its production. However, precise and accurate methods for quantification of hydroxyl radicals have not been developed so far. For this reason, this study was initiated to compare existing methods for analysis of hydroxyl radicals produced by $TiO_2$ photo-catalytic oxidation and to propose a new method to overcome the limitation of established methods. To simulate $TiO_2$ photo-catalytic oxidation process, Degussa P25 which has been widely used as a standard $TiO_2$ photo-catalyst was used with the dose of 0.05 g/L. The light source of process was UVC mercury low-pressure lamp (11 W, $2,975mW/cm^2$). The results indicate that both potassium iodide (KI)/UV-vis spectrometer and terephthalic acid (TPA)/fluorescence spectrometer methods could be applied to qualitatively measure hydroxyl radicals via detection of triiodide ion ($I_3{^-}$) and 2-hydroxyterephthalic acid which are produced by reactions of iodine ion ($I^-$) and TPA with hydroxyl radicals, respectively. However, it was possible to quantitatively measure hydroxyl radicals using TPA method coupled with high-performance liquid chromatograph (HPLC). The analytical results using TPA/HPLC method show that hydroxyl radical of 0.013 M was produced after 8 hours operation of photo-catalytic oxidation under specific experimental conditions of this study. The proposed method is expected to contribute to precise the evaluation of the performance of photo-catalytic oxidation process.

Bioequivalence of Carvelol Tablet to Dilatrend Tablet (Carvedilol 25 mg) (딜라트렌 정(카르베딜롤 25 mg)에 대한 카베롤 정의 생물학적 동등성)

  • Cho, Hea-Young;Lee, Moon-Seok;Park, Soon-Cheol;Lim, Dong-Koo;Moon, Jai-Dong;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
    • /
    • v.31 no.4
    • /
    • pp.289-295
    • /
    • 2001
  • Carvedilol is an antihypertensive and antianginal compound that combines nonselective beta-adrenoceptor blocking and vasodilation properties and is devoid of intrinsic sympathomimetic activity. The purpose of the present study was to evaluate the bioequivalence of two carvedilol tablets, $Dilatrend^{TM}$ (Chong Kun Dang Pharmaceutical Co., Ltd.) and $Carvelol^{TM}$ (Dae Won Pharmaceutical Co., Ltd.), according to the prior and revised guidelines of Korea Food and Drug Administration (KFDA). The carvedilol release from the two carvedilol tablets in vitro was tested using KP VII Apparatus II method with various different kinds of dissolution media (pH 1.2, 4.0, 6.8 buffer solution, water and blend of PSB80 into water). Eighteen normal male volunteers, $24.22{\pm}1.86$ years in age and $64.81{\pm}4.56\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After one tablet containing 25 mg of carvedilol was orally administered, blood was taken at predetermined time intervals and the concentrations of carvedilol in serum were determined using HPLC method with fluorescence detector. The dissolution profiles of two carvedilol tablets were very similar at all dissolution media. Besides, the pharmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using non-transformed and logarithmically transformed $AUC_t$ and $C_{max}$. The results showed that the differences in $AUC_t$, $C_{max}$ and $T_{max}$ between two tablets based on the $Dilatrend^{TM}$ were 2.23%, -2.00% and 0.00%, respectively. Minimum detectable differences $({\Delta})$ at ${\alpha}=0.05$ and $1-{\beta}=0.8$ were less than 20% (e.g., 13.55% and 17.61% for $AUC_t$ and $C_{max}$, respectively). The powers $(l-{\beta})$ at ${\alpha}=0.05$, ${\Delta}=0.2$ for $AUC_t$ and $C_{max}$ were 98.08% and 88.81%, respectively. The 90% confidence intervals were within ${\pm}20%$ (e.g., $-5.69{\sim}10.16$ and $-12.30{\sim}8.30$ for $AUC_t$ and $C_{max}$, respectively). There were no sequence effect between two tablets in logarithmically transformed $AUC_t$ and $C_{max}$. The 90% confidence intervals using logarithmically transformed were within the acceptance range of log(0.8) to log(1.25) (e.g., $0.95{\sim}1.11$ and $0.89{\sim}1.09$ for $AUC_t$ and $C_{max}$, respectively). Two parameters met the criteria of prior and revised KFDA guideline for bioequivalence, indicating that $Carvelol^{TM}$ tablet is bioequivalent to $Dilatrend^{TM}$ tablet.

  • PDF

Analysis Corrosion Products Formed on the Great Buddha Image of Kotokuin Temple in Kamakura (고덕원 국보 동조아미타여래좌상의 표면에 생성한 부식생성물의 해석)

  • Matsuda Shiro;Aoki Shigeo;Kang, Dai-il
    • 보존과학연구
    • /
    • s.17
    • /
    • pp.161-182
    • /
    • 1996
  • In natural atmosphere, copper and copper alloy have been used to make buddha statues and ornaments of historic buildings since the abovementioned metals have corrosion resistance in some extent, and the patinaformed on the surface of the metals has provided the people aesthetic satisfaction with its beauty. But in atmosphere polluted by $SO_x$and $NO_x$, the patina layer does not work as a protective film, and it allows damages of the metal. Since 1992, Tokyo National Research Institute of Cultural Properties(TNRICP)has conducted studies on the influence of atmospheric pollution on metal cultural property held under open air. The Great Buddha Image which is located in Kamakura about 50km west from Tokyo, has been selected as one of the objects to study because it is made by copper alloy and it has stood exposed in the air for about a few hundreds years. Furthermore it is also the reason to study on it that there are many cultural properties in the surroundings of it. We have analysed the components and the structure of the corrosion products formed on the surface of the Buddha, have carried out exposure tests using the alloy samples which have simulated the components of the Great Image, and have observed climated and polluted air in order to discuss the relation between corrosion of metals in open air and conditions of the atmosphere. In this paper, the authors have described the components and the structure of the corrosion product formed on the surface of the Great Image by means of X-ray fluorescence spectroscopy and X-ray diffraction. The conclusions are as follows. (1) Sulfate patina composed mainly with brochantite were detected on the all sides of the Image and the amount of the patina is found more on the back of the Image facing to north. (2) Antlerite were detected on the back and a park of the left side facing to west, and formation of it was considered to have close relation with malignant atmosphere. (3) A big amount of chloride patina which mainly composed of atacamite were observed on the front facing to south. (4) Carbonate patina mainly composed of malachite were detected on the area where brochantite was often detected as well. It suggested that malachite had been transformed into brochantite by deteriorated atmosphere. (5) On the all sides of the Image, patina were observed together with copper oxides mainly composed of cuprous oxide. It showed that the surface layer of the Image consists of two layers : inner layer of oxide and outer layer of patina. (6) Corrosion products of lead which was a component of copperalloy were detected on the all sides : the main lead product found on the front was chlorophosphate whereas the one on the back was sulfate.

  • PDF

A Study on Contents of Vitamin K1 in Local Agricultural Products (지역농산물의 비타민 K1 함량 조사)

  • Lee, A Reum;Kim, Joo Hee;Park, Jae-Ho;Kim, Youngho;Hong, Eui Yon;Kim, Haeng-Ran;Choi, Youngmin;Lee, Junsoo;Eom, Hyun-Ju
    • The Korean Journal of Food And Nutrition
    • /
    • v.29 no.3
    • /
    • pp.301-306
    • /
    • 2016
  • Vitamin K (phylloquinone) is an essential cofactor in the synthesis of active blood-clotting factors II, VII, IX and X. Deficiency of vitamin K leads to inadequate activity of these factors, resulting in bleeding. In this study, we investigated vitamin $K_1$ content of agricultural products that are widely and specifically grown in Korea including 9 leaves and vegetables, 16 fruits, and 11 cereals and specialty crops. Vitamin $K_1$ analysis of the agro-samples was by a validated, modified, reversed phase-HPLC method with fluorescence detection after post-column derivatization. The vitamin $K_1$ content ranged from 1.83 to $682.73{\mu}g$/100 g in leaves and vegetables, 0.17 to $28.22{\mu}g$/100 g in fruits, and ND to $279{\mu}g$/100 g in cereals and specialty crops. Among the 36 samples, high content of vitamin $K_1$ were found in Gugija (Lycium chinense Miller) leaves (average $682.73{\mu}g$/100 g) and Hansan ramie leaves (average $423.12{\mu}g$/100 g); however, mushroom, amaranth and Chinese artichoke showed no detectable levels. The results of ourstudy provide reliable vitamin $K_1$ content of Korean grown agricultural products that expand nutritional information and food composition database.

Studies on the Control of the Pine Bark Beetle (Myelophilus pinipedera LINNE) (I) -Analysis of the carotenoids in important pine species in Korea- (소나무좀 방제(防除)에 관한 연구(硏究)(제1보(第1報)) -한국산(韓國産) 중요(重要)소나무류(類) 잎에 함유(含有)되어 있는 Carotenoid 분석(分析)-)

  • Kim, Young Ho
    • Journal of Korean Society of Forest Science
    • /
    • v.27 no.1
    • /
    • pp.15-21
    • /
    • 1975
  • According to Yoshihisa Kuroki's report that the total amount of carotenoid was more in the susceptible to pine bark beetle, than in the resistant ones, carotenoids were extracted from needle leaves of one year old seedlings of Pinus thunbergii, Pinus desiflora, Pinus rigida, Pinus koraiensis, Pinus rigitaeda and Pinus taeda which are all important pines in Korea, to find their resistant ratio to the insect. The carotenoids were analyzed and compared using the spectra of them by spectrophotometer. The results were as follows: 1. The visible absorption spectra of carotenoids in those pine trees were proved to be very similar. 2. The total amount of carotenoids in needles differed with the tree species and the contents were arranged in decreasing order P. koraiensis>P. rigida>P. thunbergii>P. rigitaeda>P. taeda, it seemed that there was corelation between the cartenoid content and the extent of harm caused by the pine bark beetles except P. koraiensis. 3. But carotenoids were contained in Robinia pseudoacacia, Castanea crenata, Chamaecyparis obtusa and Cedrus deodra leaves too. 4. The total amounts of carotenoids in these pines of 9 species were arranged in decreasing order Robinia pseudoacacia>Pinus koraiensis>Pinus rigida>Pinus thunbergii>Castanea crenata>Pinus rigitaeda>Pinus taeda>Chamaecyparis obtusa>Cedrus deodara. Therefore, it was proved that there was no correlation between carotenoid cotent and extent of resistance to the insect. 5. In the thin-layer chromatography of these carotenoids, 13 kinds of components in P. densiftora, P. koraiensis and P. rigida and 12 kinds of spots in other pines, were detected respectively, under ultra-violet fluorescent lamp $3,600{\AA}$ and $2,537{\AA}$. 6. The eighth spots from the bottom in P. densiflora, P. koraiensis and P. rigida were not found in other pines and other 4 species (Robinia pseudoacacia, Castanea crenata, Chamacyparis obtusa, Cedrus deodra). Especially the spot in P. densiflora fluoresced strong cobalt blue-fluorescence under ultra-violet fluorescent lamp $2,537{\AA}$.

  • PDF

Isolation and characterization of human dental tissue-derived stem cells in the impacted wisdom teeth: comparison of dental follicle, dental pulp, and root apical papilla-derived cells (미성숙 매복지치의 치낭, 치수, 치근유두 조직에서 다능성 줄기세포의 분리와 특성화에 대한 연구)

  • Song, Jung-Ho;Park, Bong-Wook;Byun, June-Ho;Kang, Eun-Ju;Rho, Gyu-Jin;Shin, Sang-Hun;Kim, Uk-Kyu;Kim, Jong-Ryoul
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
    • /
    • v.36 no.3
    • /
    • pp.186-196
    • /
    • 2010
  • Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.

Comparison of Stress Response in Diallel Crossed Korean Domestic Chicken Breeds (토종 종계를 이용한 이면 교배조합 계통 간 스트레스 반응정도 비교 분석)

  • Cho, Eun Jung;Park, Ji Ae;Choi, Eun Sik;Sohn, Sea Hwan
    • Korean Journal of Poultry Science
    • /
    • v.43 no.2
    • /
    • pp.77-88
    • /
    • 2016
  • To establish a new synthetic Korean meat chicken breed, we tested $5{\times}5$ diallel cross mating experiment with domestic chicken breeds. Comparing stress responses among diallel crossed chicken breeds, we analyzed telomere length, DNA damage and expressions of heat shock protein genes (HSPs) as the markers of the stress response. The telomere length was measured by quantitative fluorescence in situ hybridization on the nuclei of lymphocytes. The expression levels of HSP-70, $HSP-90{\alpha}$ and $HSP-90{\beta}$ genes were analyzed by quantitative real-time polymerase chain reaction in lymphocytes. The DNA damage rate of lymphocytes was quantified by the comet assay known as the single cell gel electrophoresis. In results, there were significant differences in the values of the stress markers such as telomere length, HSPs and DNA damage rate, and also were significant differences in viabilities and body weights among the $5{\times}5$ diallel crossed chicken breeds. The telomere shortening rate, expression values of HSPs and DNA damage rate were significant low in W and Y crossed chickens compare to the others, but GG pure breed showed the highest values in the 25 crossed chickens. Estimating correlation coefficient, the survival rate positively correlated to telomere length, but negatively correlated to the expression levels of HSP-70, $HSP-90{\alpha}$, $HSP-90{\beta}$ genes and to the value of % DNA in tail as DNA damage rate. The expression levels of HSP-70, $HSP-90{\alpha}$ and $HSP-90{\beta}$ genes of dead chickens had significantly higher than those of survival chickens. According to the results on the stress marker analysis, it would be considered that the crossed breeds had more stress resistant than the pure breeds, and the crossed chickens with a light strain such as W or Y were relatively resistant to stress, but the crossed chickens with a heavy strain such as G, H, F were susceptible to stress.

Analysis and Risk Assessment of Benzo(a)pyrene in Edible Oils (식용유지류 중 벤조피렌의 함량 분석 및 안전성 평가)

  • Jang, Mi-Ra;Hong, Mi-Sun;Jung, So-Young;Choi, Bu-Chuhl;Lee, Kyeong-Ah;Kum, Jin-Young;Kim, Il-Young;Kim, Jung-Hun;Chae, Young-Zoo
    • Journal of Food Hygiene and Safety
    • /
    • v.29 no.2
    • /
    • pp.141-145
    • /
    • 2014
  • To assess the health risk for benzo(a)pyrene by the intake of edible oils, 288 cases of edible oils collected from food markets were analysed using the high performance liquid chromatography with fluorescence detector. The levels of benzo(a)pyrene were from non-detection to $4.78{\mu}g/kg$, and the average was $0.11{\mu}g/kg$. The chronic daily exposures of benzo(a)pyrene for total population group and consumer-only group were estimated using the food consumption data in the fifth Korea National Health and Nutrition Examination Survey in 2011. The estimated daily intake of benzo(a)pyrene was $4.26{\times}10^{-3}ng/kg$ b.w./day for total population group and $7.64{\times}10^{-3}ng/kg$ b.w./day for consumer-only group. The MOE (margin of exposure) of benzo(a)pyrene for total population group and consumer-only group was $7.28{\times}10^7{\sim}1.74{\times}10^8$ and $3.95{\times}10^7{\sim}9.42{\times}10^7$, respectively. Accordingly, the health risk from benzo(a)pyrene caused by the intake of edible oils was considered as a very low level.

Determination of Polycyclic Aromatic Hydrocarbons in Smoked Food Products (훈연식품 중 polycyclic aromatic hydrocarbons 함량 분석)

  • Seo, Ilwon;Nam, Hejung;Lee, Songyoung;Lee, Kyueun;Shin, Han-Seung
    • Food Engineering Progress
    • /
    • v.13 no.3
    • /
    • pp.195-202
    • /
    • 2009
  • This study was accomplished that analysis of seven polycyclic aromatic hydrocarbons (PAHs) in smoked or nonsmoked processing foods by high performance liquid chromatography (HPLC) with fluorescence detection. The calibration line was constructed with injected different levels of standard concentration. Limit of detection (LOD) and limit of quantification(LOQ) showed higher linearity ($r^{2}$=0.998) reasonably, and recovery exhibited 0.033-0.666 $\mu$g/kg, 0.108-2.217 $\mu$g/kg and 69.31-90.14%, respectively. As a result, the samples using smoked tuna as smoked materials contained seven PAHs with different range from 0.256 to 0.486 $\mu$g/kg. The benzo[a]pyrene, indicator of PAHs, was detected to below the LOQ in two samples. Concentrations of benzo[a]pyrene in three samples were below the 2 $\mu$g/kg which is the limit of regulation. Smoked tuna sauces were detected from 0.321 to 0.552 $\mu$g/kg and not detected in drying powders. PAHs of smoked meat products were ranged from 0.720 to 2.027 $\mu$g/kg and are higher than concentration of tuna smoked samples. PAHs were very low in non-smoked foods including mustard, herb, and roasted meats.